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. 1987 Jul;25(7):1269–1274. doi: 10.1128/jcm.25.7.1269-1274.1987

Genetic relatedness among human rotavirus genes coding for VP7, a major neutralization protein, and its application to serotype identification.

K Midthun, J Flores, K Taniguchi, S Urasawa, A Z Kapikian, R M Chanock
PMCID: PMC269191  PMID: 3038948

Abstract

Antigenic characterization of human rotaviruses by plaque reduction neutralization assay has revealed four distinct serotypes. The outer capsid protein VP7, coded for by gene 8 or 9, is a major neutralization protein; however, studies of rotaviruses derived from genetic reassortment between two strains have confirmed that another outer capsid protein, VP3, is in some cases equally important in neutralization. In this study, the genetic relatedness of the genes coding for VP7 of human rotaviruses belonging to serotypes 1 through 4 was examined by hybridization of their denatured double-stranded genomic RNAs to labeled single-stranded mRNA probes derived from human-animal rotavirus reassortants containing only the VP7 gene of their human rotavirus parent. A high degree of homology was demonstrated between the VP7 genes of strain D and other serotype 1 human rotaviruses, strain DS-1 and other serotype 2 human rotaviruses, strain P and other serotype 3 human rotaviruses, and strain ST3 and other serotype 4 human rotaviruses. Hybrid bands could not be demonstrated between the VP7 gene of D, DS-1, P, or ST3 and the corresponding gene of human rotaviruses belonging to a different serotype. RNA specimens extracted from the stools of 15 Venezuelan children hospitalized with rotavirus diarrhea were hybridized to each of the reassortant probes representing the four human serotypes. All five viruses with short RNA patterns showed homology with the DS-1 strain VP7 gene; two of these were previously adapted to tissue culture and shown to be serotype 2 strains by tissue culture neutralization. Of the remaining 10 viruses with long RNA patterns, 2 hybridized only to the D strain VP7 gene, 6 hybridized only to the P strain VP7 gene, and 2 hybridized only to the ST3 strain VP7 gene. Hybridization using single human rotavirus gene substitution reassortants as probes may provide an alternative method for identifying the VP7 serotype of field isolates that would circumvent the need for tissue culture adaptation.

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Selected References

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  1. Albert M. J., Bishop R. F., Shann F. A. Epidemiology of rotavirus diarrhea in the Highlands of Papua, New Guinea, in 1979, as revealed by electrophoresis of genome RNA. J Clin Microbiol. 1983 Jan;17(1):162–164. doi: 10.1128/jcm.17.1.162-164.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beards G. M., Flewett T. H. Serological characterisation of human rotaviruses propagated in cell cultures. Brief report. Arch Virol. 1984;80(2-3):231–237. doi: 10.1007/BF01310663. [DOI] [PubMed] [Google Scholar]
  3. Beards G. M., Pilfold J. N., Thouless M. E., Flewett T. H. Rotavirus serotypes by serum neutralisation. J Med Virol. 1980;5(3):231–237. doi: 10.1002/jmv.1890050307. [DOI] [PubMed] [Google Scholar]
  4. Estes M. K., Graham D. Y., Dimitrov D. H. The molecular epidemiology of rotavirus gastroenteritis. Prog Med Virol. 1984;29:1–22. [PubMed] [Google Scholar]
  5. Flores J., Greenberg H. B., Myslinski J., Kalica A. R., Wyatt R. G., Kapikian A. Z., Chanock R. M. Use of transcription probes for genotyping rotavirus reassortants. Virology. 1982 Sep;121(2):288–295. doi: 10.1016/0042-6822(82)90168-4. [DOI] [PubMed] [Google Scholar]
  6. Flores J., Perez-Schael I., Boeggeman E., White L., Perez M., Purcell R., Hoshino Y., Midthun K., Chanock R. M., Kapikian A. Z. Genetic relatedness among human rotaviruses. J Med Virol. 1985 Oct;17(2):135–143. doi: 10.1002/jmv.1890170206. [DOI] [PubMed] [Google Scholar]
  7. Flores J., Perez I., White L., Perez M., Kalica A. R., Marquina R., Wyatt R. G., Kapikian A. Z., Chanock R. M. Genetic relatedness among human rotaviruses as determined by RNA hybridization. Infect Immun. 1982 Aug;37(2):648–655. doi: 10.1128/iai.37.2.648-655.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gerna G., Passarani N., Battaglía M., Percivalle E. Rapid serotyping of human rotavirus strains by solid-phase immune electron microscopy. J Clin Microbiol. 1984 Feb;19(2):273–278. doi: 10.1128/jcm.19.2.273-278.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Glass R. I., Keith J., Nakagomi O., Nakagomi T., Askaa J., Kapikian A. Z., Chanock R. M., Flores J. Nucleotide sequence of the structural glycoprotein VP7 gene of Nebraska calf diarrhea virus rotavirus: comparison with homologous genes from four strains of human and animal rotaviruses. Virology. 1985 Mar;141(2):292–298. doi: 10.1016/0042-6822(85)90260-0. [DOI] [PubMed] [Google Scholar]
  10. Greenberg H. B., Flores J., Kalica A. R., Wyatt R. G., Jones R. Gene coding assignments for growth restriction, neutralization and subgroup specificities of the W and DS-1 strains of human rotavirus. J Gen Virol. 1983 Feb;64(Pt 2):313–320. doi: 10.1099/0022-1317-64-2-313. [DOI] [PubMed] [Google Scholar]
  11. Gunn P. R., Sato F., Powell K. F., Bellamy A. R., Napier J. R., Harding D. R., Hancock W. S., Siegman L. J., Both G. W. Rotavirus neutralizing protein VP7: antigenic determinants investigated by sequence analysis and peptide synthesis. J Virol. 1985 Jun;54(3):791–797. doi: 10.1128/jvi.54.3.791-797.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hoshino Y., Sereno M. M., Midthun K., Flores J., Kapikian A. Z., Chanock R. M. Independent segregation of two antigenic specificities (VP3 and VP7) involved in neutralization of rotavirus infectivity. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8701–8704. doi: 10.1073/pnas.82.24.8701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hoshino Y., Wyatt R. G., Flores J., Midthun K., Kapikian A. Z. Serotypic characterization of rotaviruses derived from asymptomatic human neonatal infections. J Clin Microbiol. 1985 Mar;21(3):425–430. doi: 10.1128/jcm.21.3.425-430.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hoshino Y., Wyatt R. G., Greenberg H. B., Flores J., Kapikian A. Z. Serotypic similarity and diversity of rotaviruses of mammalian and avian origin as studied by plaque-reduction neutralization. J Infect Dis. 1984 May;149(5):694–702. doi: 10.1093/infdis/149.5.694. [DOI] [PubMed] [Google Scholar]
  15. Kalica A. R., Greenberg H. B., Espejo R. T., Flores J., Wyatt R. G., Kapikian A. Z., Chanock R. M. Distinctive ribonucleic acid patterns of human rotavirus subgroups 1 and 2. Infect Immun. 1981 Sep;33(3):958–961. doi: 10.1128/iai.33.3.958-961.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kalica A. R., Greenberg H. B., Wyatt R. G., Flores J., Sereno M. M., Kapikian A. Z., Chanock R. M. Genes of human (strain Wa) and bovine (strain UK) rotaviruses that code for neutralization and subgroup antigens. Virology. 1981 Jul 30;112(2):385–390. doi: 10.1016/0042-6822(81)90285-3. [DOI] [PubMed] [Google Scholar]
  17. Kapikian A. Z., Wyatt R. G., Greenberg H. B., Kalica A. R., Kim H. W., Brandt C. D., Rodriguez W. J., Parrott R. H., Chanock R. M. Approaches to immunization of infants and young children against gastroenteritis due to rotaviruses. Rev Infect Dis. 1980 May-Jun;2(3):459–469. doi: 10.1093/clinids/2.3.459. [DOI] [PubMed] [Google Scholar]
  18. Matthews R. E. The classification and nomenclature of viruses. Summary of results of meetings of the International Committee on Taxonomy of Viruses in The Hague, September 1978. Intervirology. 1979;11(3):133–135. doi: 10.1159/000149025. [DOI] [PubMed] [Google Scholar]
  19. Midthun K., Greenberg H. B., Hoshino Y., Kapikian A. Z., Wyatt R. G., Chanock R. M. Reassortant rotaviruses as potential live rotavirus vaccine candidates. J Virol. 1985 Mar;53(3):949–954. doi: 10.1128/jvi.53.3.949-954.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Midthun K., Hoshino Y., Kapikian A. Z., Chanock R. M. Single gene substitution rotavirus reassortants containing the major neutralization protein (VP7) of human rotavirus serotype 4. J Clin Microbiol. 1986 Nov;24(5):822–826. doi: 10.1128/jcm.24.5.822-826.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Offit P. A., Blavat G. Identification of the two rotavirus genes determining neutralization specificities. J Virol. 1986 Jan;57(1):376–378. doi: 10.1128/jvi.57.1.376-378.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Perez-Schael I., Daoud G., White L., Urbina G., Daoud N., Perez M., Flores J. Rotavirus shedding by newborn children. J Med Virol. 1984;14(2):127–136. doi: 10.1002/jmv.1890140206. [DOI] [PubMed] [Google Scholar]
  23. Rodger S. M., Bishop R. F., Birch C., McLean B., Holmes I. H. Molecular epidemiology of human rotaviruses in Melbourne, Australia, from 1973 to 1979, as determined by electrophoresis of genome ribonucleic acid. J Clin Microbiol. 1981 Feb;13(2):272–278. doi: 10.1128/jcm.13.2.272-278.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rodriguez W. J., Kim H. W., Brandt C. D., Gardner M. K., Parrott R. H. Use of electrophoresis of RNA from human rotavirus to establish the identity of strains involved in outbreaks in a tertiary care nursery. J Infect Dis. 1983 Jul;148(1):34–40. doi: 10.1093/infdis/148.1.34. [DOI] [PubMed] [Google Scholar]
  25. Sato K., Inaba Y., Miura Y., Tokuhisa S., Matumoto M. Antigenic relationships between rotaviruses from different species as studied by neutralization and immunofluorescence. Arch Virol. 1982;73(1):45–50. doi: 10.1007/BF01341726. [DOI] [PubMed] [Google Scholar]
  26. Shaw R. D., Stoner-Ma D. L., Estes M. K., Greenberg H. B. Specific enzyme-linked immunoassay for rotavirus serotypes 1 and 3. J Clin Microbiol. 1985 Aug;22(2):286–291. doi: 10.1128/jcm.22.2.286-291.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Taniguchi K., Urasawa T., Morita Y., Greenberg H. B., Urasawa S. Direct serotyping of human rotavirus in stools by an enzyme-linked immunosorbent assay using serotype 1-, 2-, 3-, and 4-specific monoclonal antibodies to VP7. J Infect Dis. 1987 Jun;155(6):1159–1166. doi: 10.1093/infdis/155.6.1159. [DOI] [PubMed] [Google Scholar]
  28. Thouless M. E., Beards G. M., Flewett T. H. Serotyping and subgrouping of rotavirus strains by the ELISA test. Arch Virol. 1982;73(3-4):219–230. doi: 10.1007/BF01318076. [DOI] [PubMed] [Google Scholar]
  29. Urasawa S., Urasawa T., Taniguchi K. Three human rotavirus serotypes demonstrated by plaque neutralization of isolated strains. Infect Immun. 1982 Nov;38(2):781–784. doi: 10.1128/iai.38.2.781-784.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wyatt R. G., Greenberg H. B., James W. D., Pittman A. L., Kalica A. R., Flores J., Chanock R. M., Kapikian A. Z. Definition of human rotavirus serotypes by plaque reduction assay. Infect Immun. 1982 Jul;37(1):110–115. doi: 10.1128/iai.37.1.110-115.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wyatt R. G., James H. D., Jr, Pittman A. L., Hoshino Y., Greenberg H. B., Kalica A. R., Flores J., Kapikian A. Z. Direct isolation in cell culture of human rotaviruses and their characterization into four serotypes. J Clin Microbiol. 1983 Aug;18(2):310–317. doi: 10.1128/jcm.18.2.310-317.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]

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